JPH06237788A - Production of acetylene alcohol compounds - Google Patents

Abstract

PURPOSE:To provide a method for industrially advantageously producing an optically active acetylene alcohol compound. CONSTITUTION:The method for producing an acetylene alcohol such as optically active 6-heptyne-2-ol comprises reacting an acetylene alcohol such as 6-heptyne-2- ol with an unsaturated alcohol in the presence of an esterase.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a process for producing acetylene alcohols to obtain compounds useful as intermediates for pharmaceuticals, agricultural chemicals and liquid crystals.

[0002]

2. Description of the Related Art As a method for synthesizing optically active acetylene alcohols according to the present invention, Japanese Patent Laid-Open No. 3-201996
The following method is described in the publication. The above-mentioned conventional method is one in which a hydrolase is reacted with an alcohol-organic solvent to carry out alcoholysis, and although a relatively high optical purity has been achieved in the obtained ester, the hydrolyzed alcohol has an optical purity. There is no description of purity etc. According to this method, the ester is further chemically hydrolyzed to obtain an alcohol having a relatively high optical purity. In general, in order for the ester product which is a hydrolysis residue to have a high optical activity purity, it is necessary to increase the hydrolysis rate, and as a result, the optical purity of the alcohol becomes insufficient.

[0003]

DISCLOSURE OF THE INVENTION Problems to be Solved by the Invention
The present inventors have earnestly studied how to obtain any enantiomer of acetylene alcohol with good optical purity and high yield, and as a result, racemic acetylene alcohols were esterified with carboxylic acid ester of unsaturated alcohol to be acylated. When reacted as an agent, an enantiomer of the optically active acetylene alcohol represented by the above formula and an optically active acetylene alcohol ester derivative (this ester derivative is also an enantiomer) are obtained. The present invention has been completed by finding that acetylene alcohols having the same configuration as the above formula and higher optical purity can be obtained in good yield if hydrolyzed.

[0004]

That is, the present invention has the general formula [1] (In the formula, n represents an integer of 0 to 6) and an carboxylic acid ester of an unsaturated alcohol as an acylating agent are reacted in the presence of esterase to give a compound represented by the general formula [2]. (In the formula, n represents the same meaning as described above, and * represents an asymmetric carbon), and an optically active acetylene alcohol represented by the general formula [3] (Wherein n and * have the same meanings as described above, and R ¹ represents an optionally halogenated alkyl group having 1 to 5 carbon atoms), and a mixture of the optically active acetylene alcohol ester derivative. And a method for producing the optically active acetylene alcohol represented by the general formula [2] and the optically active acetylene alcohol ester derivative represented by the general formula [3], characterized by isolating them, and the optically active acetylene alcohol ester. The present invention relates to a method for asymmetrically hydrolyzing a derivative using esterase to obtain an enantiomer of the optically active acetylene alcohol represented by the general formula [2].

The present invention will be described in detail below. In the present invention, examples of the acetylene alcohol represented by the general formula [1] that is a raw material include 3-butyn-2-ol, 4-pentyn-2-ol, 5-hexyn-2-ol, 6-heptin- 2-ol, 7-octin-2-ol, 8-nonin-2-ol, 9-decin-2-ol can be exemplified.

Examples of the acylating agent to be used include carboxylate esters of unsaturated alcohols such as vinyl acetate, vinyl propionate, vinyl valerate, isopropenyl acetate, isopropenyl propionate and isopropenyl valerate. Or a halogenated acyl ester thereof.

The amount used is usually 0.5 mol times or more, preferably 2 mol times or more, with respect to the acetylene alcohol [1]. Also, an acylating agent can be used as a solvent.

The esterase having the ability to selectively esterify only one of the optical isomers of acetylene alcohols in this reaction means a broadly defined esterase containing lipase. Among them, lipase is preferable, and more specifically, lipase derived from Arthrobacter genus and Pseudomonas genus can be mentioned.

In this reaction, an enzyme obtained from an animal, a plant or a microorganism is used as the esterase, and the usage forms thereof include a purified enzyme, a crude enzyme, an enzyme-containing material, a microbial culture solution, a culture, a microbial cell, It can be used in various forms such as a culture filtrate or a treated product thereof, if necessary, and an enzyme and a microorganism can be used in combination. Alternatively, it can also be used as an enzyme immobilized on a resin or the like, or an immobilized cell.

The microorganisms that produce esterase include
There is no particular limitation as long as it is a microorganism that produces an esterase having the ability to selectively esterify only one of the optical isomers of acetylene alcohols.
Specific examples of such microorganisms include, for example, Enterobacter, Arthrobacter, Brevibacterium, Pseudomonas, Alcaligenas, Micrococcus, Bacillus, Lactobacillus, Trichoderma, Candida, Saccharomyces. , Rhodotorula, Cryptococcus, Tollopsis, Pichia,
Examples include microorganisms belonging to the genus Penicillium, Aspergillus, Rhizopus, Mucor, Aureopasidium, Actinomucor, Nocardia, Streptomyces, Hansenula, and Achromobacter.

Cultivation of the above-mentioned microorganisms is usually carried out according to a conventional method, and for example, liquid culture can be carried out to obtain a culture solution. For example, sterilized liquid medium [molds,
Malt extract / yeast extract medium for yeasts (dissolve 5 g of peptone, 10 g of glucose, 3 g of malt extract, 3 g of yeast extract to pH 6.5 in 1 L of water), and a sweetened broth medium for bacteria (1 L of water) Peptone 5g, glucose 10
g, meat extract 3 g, and NaCl 3 g to adjust the pH to 7.2)] and inoculate with a microorganism, and usually cultivated at 20 to 40 ° C for 1 to 3 days, and if necessary, solid culture is performed. You can go.

Some of these esterases of microbial origin are commercially available and can be easily obtained. Specific examples of commercially available esterases include, for example, Pseudomonas lipase [Lipase Amano PS, Lipase Amano P (both manufactured by Amano Pharmaceuticals)], Aspergillus lipase [Lipase Amano AP, Lipase Amano A1].
2 (both manufactured by Amano Pharmaceuticals)], lipase of Mucor genus [Lipase Amano M-AP (manufactured by Amano Pharmaceuticals)], Candida
Cylindrace lipase [Lipase MY (manufactured by Meito Sangyo)], Alcaligenes lipase [Lipase PL
(Manufactured by Meito Sangyo)], lipase of Achromobacter genus [Lipase AL (manufactured by Meito Sangyo)], lipase of genus Arthrobacter [manufactured by Shin Nihon Kagaku], lipase of genus Chromobacterium (manufactured by Toyo Brewery) Examples thereof include Rhizopus derema lipase [Taripase (manufactured by Tanabe Seiyaku)], Rhizopus lipase [Lipase Saiken (Osaka Bacterial Research Institute)], and the like.

Animal / plant esterases can also be used. Specific examples of these esterases include steapsin, pancreatin, pig liver esterase,
Wheat Germ esterase etc. can be mentioned. The amount of enzyme is usually 0. 0 for acetylene alcohols [1].
It is 001 to equivalent times, preferably 0.003 to 0.5 times. Of course, more than this amount may be used.

A solvent may be used in this reaction, and as the solvent, other than the above-mentioned acylating agent, for example, hexane, heptane, benzene, acetone, toluene,
There may be mentioned aliphatic or aromatic hydrocarbons such as dichloromethane, dibutyl ether, etc., halogenated hydrocarbons, ethers, ketones etc. alone or in a mixture. The amount used is acetylene alcohols [1]
On the other hand, it is usually 0.5 to 10 times by weight. Of course, there is no problem in the reaction even if an amount exceeding 10 times by weight is used.

The reaction temperature is usually 10 to 50 ° C., and the reaction time is not particularly limited, but 0.5 to 60 hours is usually sufficient.
The progress of the reaction can be traced by measuring the optical purity of the optically active acetylene alcohol [1] in the reaction system, for example, by liquid chromatography equipped with a filler for separating the optically active compound, and thereby the reaction end point. You can also decide.

After completion of the reaction, if necessary, an aliphatic or aromatic hydrocarbon such as hexane, heptane, benzene, acetone, toluene, dichloromethane, chloroform, chlorobenzene, dichloroethane, ethyl acetate, ethyl ether, ether, or ketone is added to the reaction mass. , An ester, a halogenated hydrocarbon, and the like are added, and the enzyme is filtered off, and the filtrate is concentrated to give an optically active acetylene alcohol represented by the general formula [2] and the general formula [3].
The optically active acetylene alcohol ester derivative represented by can be obtained. Further, it is also possible to separate the optically active acetylene alcohols represented by the general formula [2] and the optically active acetylene alcohol ester derivatives represented by the general formula [3] by subjecting them to a conventional chromatography treatment. . The optically active acetylene alcohol ester derivative [3] thus obtained can be converted into optically active acetylene alcohols which are antipodes by further asymmetric hydrolysis using esterase. As the esterase used in the asymmetric hydrolyzation reaction, the same esterase as the one described above can be used.

The asymmetric hydrolysis reaction is carried out by vigorously stirring the mixture of the acetylene alcohol ester derivative obtained above and the above enzyme or microorganism in a buffer solution. As the buffer solution, a commonly used buffer solution of an inorganic acid salt such as sodium phosphate and potassium phosphate, a buffer solution of an organic acid salt such as sodium acetate and sodium citrate, and the like can be used. The pH of 8-11 is preferable for the culture broth and alkaline esterase, and the pH of 5-8 is preferable for the culture broth of non-alkalophilic microorganisms and the esterase having no alkali tolerance. Concentration is usually 0.05─
It is in the range of 3M, preferably 0.05-0.5M.

The reaction temperature is usually 10 to 60 ° C., and the reaction time is generally 10 to 70 hours, but it is not limited thereto. When a lipase belonging to the genus Pseudomonas or the genus Arthrobacter is used as the lipase in this asymmetric hydrolysis reaction, the optically active acetylene alcohols of the object can be obtained with a relatively high optical purity. In addition, in the case of this asymmetric hydrolysis reaction, in addition to the buffer solution, it is also possible to use an organic solvent inert to the reaction such as toluene, chloroform, methyl isobutyl ketone, dichloromethane, etc. It can be done in an advantageous manner.

After completion of such hydrolysis reaction, the hydrolysis reaction solution is subjected to extraction treatment with a solvent such as ethyl ether or dichloromethane, the solvent is distilled off from the organic layer, and the concentrated residue is treated with column chromatography. By the method described above, the optically active acetylene alcohols, which are hydrolysis products, can be separated.

[0020]

Industrial Applicability According to the method of the present invention, any enantiomer of an optically active acetylene alcohol can be obtained in a short time with a high optical yield and a high yield, which is industrially advantageous.

[0021]

The present invention will be described in more detail with reference to the following examples, but the present invention is not limited to the examples.

Example 1 1-butyn-3-ol [1
-1] 10 g, Lipase Amano PS (Amano Pharmaceutical Co., Ltd.) 0.2 g and
Stir the mixture of vinyl acetate and 50 ml of vinyl acetate at 35 ° C and stir.
did. On the way, check the reaction solution by gas chromatography.
I clicked. The reaction was stopped 10 hours after the start of stirring, and the lipase was added.
Was removed by filtration, the filter residue was washed with ethyl ether and filtered,
The solution was concentrated under reduced pressure together with the above filtrate. The obtained concentrate
Column chromatography separation and purification, (S)-(-)-1-butyne-3
-All [2-1] 3.5 g (optical purity 86% e.e., [α]_D
²⁰-51 ° (neat) and R rich-1-butyn-3-o
To obtain 10.2 g of the acetic acid ester [3-1] of the alcohol.
The optical purity of the compound is dinitrophenyl isocyanate.
Liquid column using optically active column after treatment
It was measured by graphy. Then I got here (3-1)
4.48g, Lipase Amano PS (Amano Pharmaceutical Co., Ltd.) 0.13g, 0.2
M phosphate buffer (pH 6.5) 60 ml mixed solution
Is stirred vigorously at 30 ° C. for 15 hours. The resulting mixture
Was extracted with 40 ml of ethyl ether, and the organic layer was washed with water.
After that, concentrate. The obtained residue is silica gel column chromatography.
Chromatography (eluent: dichloromethane-hexane)
Release, R (+)-1-butyn-3-ol (4-1) 1.
55 g (optical purity 94.6%) [α]_D ²⁰+ 57 ° (nea
get t).

Example 2 The same operation as in Example 1 was carried out by substituting vinyl butyrate for vinyl acetate. After agitation 18
The reaction was stopped at a certain time, and after the same post-treatment, (S) -1-butyn-3-ol [2-2] 4.1 g (optical purity 89% ee)
And R-rich-1-butyn-3-ol butyrate
11.8 g [3-2] was obtained respectively. Next, (3-2) 7.0 g, Arthrobacter lipase (Nippon Kagaku) 0.14 g, 0.2 M
A mixture of 40 ml of phosphate buffer is vigorously stirred for 10 hours at 25 ° C. The resulting mixture is ethyl ether 4
It is extracted with 0 ml, and the organic layer is washed with water and then concentrated. The obtained residue was subjected to silica gel column chromatography (eluent:
Separated with dichloromethane-hexane) and R (+)-1-
Butin-3-ol (4-1) 2.05 g (optical purity 96.4
%) [Α] _D ²⁰ + 59 ° (neat) is obtained.

Example 3 4-Pentin-2-ol
[1-3] 10 g, Lipase Amano PS (Amano Pharmaceutical Co., Ltd.) 0.2 g
Keep the mixture of 50 ml of vinyl acetate and vinyl acetate at 35 ° C and stir.
I spilled. On the way, the reaction solution was subjected to gas chromatography.
Checked. The reaction was stopped 14 hours after the start of stirring, and the
Filtered off and the filter residue washed with ethyl ether and filtered
Then, it was concentrated under reduced pressure together with the above filtrate. Obtained concentration
The product was separated and purified by column chromatography to obtain (S)-(-)-4-pliers.
N-2-ol [2-3] 3.2 g (optical purity 84.6% e.e.)
[Α]_D ²⁰-30 ° (c = 5, benzene) and (R) -4-
Pentin-2-ol acetate ester [3-3] 9.9 g
Got each. Next, (R) -4-pentyne acetate
[3-3] 3.78 g and Lipase Amano PS (Amano Pharmaceutical Co., Ltd.) 0.1
g and 50 ml of 0.3 M phosphate buffer (pH 7.0)
It was kept at 35 ° C and stirred. On the way, the reaction solution was liquid chromatographed.
I checked it with a graph. 10 hours after the start of stirring
Stop the reaction, remove the lipase by filtration, and remove the filter residue with ethyl ether.
Washed with water, extracted with ethyl ether, and the organic layer is sulfuric acid.
After drying over magnesium, it was concentrated under reduced pressure. Got
Column chromatographic separation and purification of the concentrate (R) -4-pentyne
2-ol [3-4] 1.43 g (optical purity 91.2% e.e.)
Obtained. [Α]_D ²⁰+ 33 ° (c = 5, benzene)

Example 4 A mixed solution of 10 g of 6-heptin-2-ol [1-4], 0.15 g of Arthrobacter lipase (manufactured by Shin Nihon Kagaku), and 40 ml of vinyl acetate was added.
The mixture was kept at 30 ° C and stirred. On the way, the reaction solution was checked by gas chromatography. The reaction was stopped 8 hours after the start of stirring, the lipase was filtered off, the filter residue was washed with ethyl ether and filtered, and the filtrate was concentrated under reduced pressure. The obtained concentrate is separated and purified by column chromatography,
(S)-(+)-6-Heptin-2-ol [2-4] 4.1 g (optical purity 91% ee) [α] _D ²⁰ + 13.5 ° (c = 5, benzene)
And R rich-6-heptin-2-ol acetic acid ester [3-4] 10.2 g were obtained. Then got here [
3-4] 7.7 g, Arthrobacter lipase (Nippon Kagaku) 0.15 g, 0.2 M phosphate buffer (pH 6.0) 40
The ml mixture is stirred vigorously at 30 ° C. for 15 hours. The obtained mixture is extracted with 40 ml of ethyl ether, and the organic layer is washed with water and then concentrated. The obtained residue was separated by silica gel column chromatography (eluent: dichloromethane-hexane), and R (-)-6-heptin-2-ol (4-4) 3.23 g (optical purity 97.2%) [ α] _D
²⁰ -14.8 ° (c = 5, benzene) is obtained.

(Example 5) [3-1] 4.48 obtained in Example 1
g, Arthrobacter lipase (Nippon Chemical) 0.09
A mixture of g and 30 ml of 0.2 M phosphate buffer (pH 6.0) is vigorously stirred at 30 ° C. for 11 hours. The obtained mixture is extracted with 40 ml of ethyl ether, and the organic layer is washed with water and then concentrated. The obtained residue was separated by silica gel column chromatography (eluent: dichloromethane-hexane), and R (+)-1-butyn-3-ol 1.64 g (optical purity 96.8%) [α] _D ²⁰ +58. To get the neat.

Reference Example 10 g of 4-pentyn-2-ol acetate ester and Lipase Amano PS (Amano Pharmaceutical Co., Ltd.) 0.2
A mixture of g and 100 ml of 0.3 M phosphate buffer was kept at 35 ° C. and stirred. On the way, the reaction solution was checked by liquid chromatography. The reaction was stopped 10 hours after the stirring was started, the lipase was filtered off, the filter residue was washed with ethyl ether, the aqueous layer was extracted and separated with ethyl ether, and the organic layer was dried over magnesium sulfate, It was concentrated under reduced pressure. The obtained concentrate was separated and purified by column chromatography to obtain (R) -4-pentyn-2-ol [3-4] 3.8 g (optical purity 62.5% ee
) Got.

Claims (2)

[Claims] 1. A general formula [1] (In the formula, n represents an integer of 0 to 6) and an carboxylic acid ester of an unsaturated alcohol as an acylating agent are reacted in the presence of esterase to give a compound represented by the general formula [2]. (In the formula, n represents the same meaning as described above, and * represents an asymmetric carbon.) And an optically active acetylene alcohol represented by the general formula [3]. (Wherein n and * have the same meanings as described above, and R ¹ represents an optionally halogenated alkyl group having 1 to 5 carbon atoms), and a mixture of the optically active acetylene alcohol ester derivative. A method for producing an optically active acetylene alcohol represented by the general formula [2] and an optically active acetylene alcohol ester derivative represented by the general formula [3], which comprises isolating them. 2. An acetylene alcohol represented by the general formula [1] and a carboxylic acid ester of an unsaturated alcohol as an acylating agent are allowed to react in the presence of an esterase, and represented by the general formula [2]. A mixture of the optically active acetylene alcohols and the optically active acetylene alcohol ester derivative represented by the general formula [3] is obtained, the optically active acetylene alcohol ester derivative represented by the general formula [3] is isolated, and then the acetylene alcohol ester derivative is isolated. A process for producing an optically active acetylene alcohol which is a target compound, wherein the compound [3] is asymmetrically hydrolyzed using an esterase.